Diamine oxidase for use in the treatment or prevention of attention deficit hyperactivity disorder (ADHD)

Abstract

The present invention refers to diamine oxidase for use in the treatment or prevention of attention deficit hyperactivity disorder (ADHD).

Claims

1. A method of treating attention deficit hyperactivity disorder (ADHD) in a human, the method comprising: testing diamine oxidase (DAO) activity level in plasma of the human; responsive to results of the testing being less than or equal to a pre-defined threshold, orally administering a dosage of 0.1-50 mg of DAO to the human; and wherein the pre-defined threshold is not greater than 80 HDU/ml.

2. The method according to claim 1, wherein the DAO is administered in at least one of tablets, capsules, and sachets.

3. The method according to claim 1, wherein the dosage is 2-20 mg.

4. The method according to claim 1, further comprising administering caffeine to the human.

5. The method according to claim 4, wherein a dosage of the administered caffeine is 1-100 mg.

6. The method according to claim 1, wherein the administered DAO is provided with gastric protection.

7. The method according to claim 1, wherein a form of the administered DAO is selected from the group consisting of free form, powder, lyophilised powder, microcapsules, nanocapsules, and liposomes.

8. The method according to claim 1, wherein the DAO is included in a composition.

9. The method according to claim 5, wherein the dosage of the administered caffeine is 5-50 mg.

10. The method according to claim 8, wherein the composition has an enteric coating that protects the DAO from gastric acidity.

Description

DETAILED DESCRIPTION OF THE INVENTION

(1) The first aspect of the present invention relates to the use of DAO for the preparation of a composition for the prevention or treatment of attention deficit hyperactivity disorder (ADHD), and to diamine oxidase (DAO) for use in the treatment or prevention of attention deficit hyperactivity disorder (ADHD), as well as to a composition comprising DAO for use in the prevention or treatment of attention deficit hyperactivity disorder (ADHD).

(2) The origin of the DAO used in the present invention can be biotechnological or from animal or plant extraction.

(3) When the DAO used is of non-plant origin, it will preferably be in the form of lyophilised powder. When the DAO used is of plant origin, it may also be in liquid form.

(4) DAO and compositions comprising DAO, to be used in the prevention or treatment of attention deficit hyperactivity disorder (ADHD), may be in the form of tablets, capsules or sachets containing DAO in free form, in powder, lyophilised powder, microcapsules, nanocapsules or liposomes of DAO with gastric protection.

(5) DAO may also be mixed with caffeine to potentiate the effects of prevention and treatment of attention deficit hyperactivity disorder. Accordingly, a composition comprising DAO and caffeine is also disclosed herein.

(6) Caffeine is an alkaloid of the xanthine group with stimulating properties that is used for the treatment of attention deficit hyperactivity disorder.

(7) The DAO content of the present invention is between 0.1 and 50 mg per unit dose, preferably between 2 and 20 mg.

(8) The caffeine content of the present invention is between 1 and 100 mg per unit dose, preferably between 5 and 50 mg.

(9) DAO or compositions comprising DAO for the prevention and treatment of attention deficit hyperactivity disorder can be taken before, after or with meals.

(10) The use of DAO or compositions comprising DAO of the present invention directly affects the blood histamine level and therefore the symptoms of attention deficit hyperactivity disorder as a consequence of accumulated histamine levels.

(11) The DAO or compositions comprising DAO of the present invention are prepared from DAO in free form, in powder, lyophilised powder, microcapsules, nanocapsules or liposomes of DAO that have an enteric coating protecting the DAO from gastric acidity, so that these various forms can be filled directly into sachets or introduced into a capsule or compressed to give rise to tablets. The enteric coating layer that coats the various forms rapidly disintegrates or dissolves in a neutral or alkaline medium.

(12) In the case of microgranules, the cores can be inert sugar-based cores or similar on which the DAO is applied, or these cores may already contain DAO mixed with other excipients. These excipients can be binders, surfactants, fillers, dispersants, alkaline additives or other pharmaceutically acceptable ingredients either alone or in a mixture. The binders can be cellulose-type such as hydroxypropyl methylcellulose, hydroxypropyl cellulose or carboxymethylcellulose sodium, polyvinylpyrrolidone, sugars, starches and other pharmaceutically acceptable substances with cohesive properties. Suitable surfactants are found in the groups of acceptable ionic or non-ionic surfactants such as, for example, sodium lauryl sulphate.

(13) Alternatively, DAO can be mixed with alkaline compounds and additionally mixed with suitable constituents to be formulated into a core material. These core materials can be produced by extrusion/spheronisation or by compression using various processing equipment.

(14) DAO can also be mixed with pharmaceutically acceptable alkaline substances such as salts of phosphoric acid and sodium, potassium, calcium, magnesium and aluminum, carbonic acid, citric acid or other suitably weak organic and inorganic acids; a co-precipitate of aluminum hydroxide/sodium bicarbonate; substances normally used in anti-acid preparations such as hydroxides of aluminum, calcium and magnesium; magnesium oxide or compound substances such as Al.sub.2O.sub.3.6MgO.CO.sub.2.12H.sub.2O, (Mg.sub.6Al.sub.2(OH).sub.16CO.sub.3.4H.sub.2O, MgO.Al.sub.2O.sub.3.2SiO.sub.2.nH.sub.2O or similar compounds; pH buffering substances such as tris(hydroxymethyl)aminomethane, basic amino acids and their salts or other pharmaceutically acceptable pH buffering substances.

(15) The enteric coating layers may contain pharmaceutically acceptable plasticisers to obtain the desired mechanical properties of flexibility and hardness. These plasticisers can be, for example, triacetin, citric acid esters, phthalic acid esters, cetyl alcohol, polyethylene glycols, polysorbates or other plasticisers.

(16) The present invention also relates to a method of treatment comprising the administration to a patient, presenting with the symptoms of attention deficit hyperactivity disorder or with the risk of suffering from it, of DAO or a composition comprising DAO according to any of the embodiments of the present invention in a therapeutically effective amount.

EXAMPLES

Example 1

(17) DAO tablets were prepared from microgranules containing 4% DAO, with the following formula:

(18) TABLE-US-00001 DAO  4 mg Mannitol 40 mg Microcrystalline cellulose 25 mg Hydroxypropyl cellulose 10 mg Corn starch 10 mg Citric acid  6 mg

(19) The microgranules were coated with hydroxypropyl methylcellulose.

(20) To make the tablets, the DAO microgranules were compressed with microcrystalline cellulose and sodium stearyl fumarate.

Example 2

(21) DAO tablets were prepared from microgranules containing 4% DAO and 10% caffeine with the following formula:

(22) TABLE-US-00002 DAO  4 mg Caffeine 10 mg Mannitol 35 mg Microcrystalline cellulose 15 mg Hydroxypropyl cellulose 10 mg Hydroxypropyl methylcellulose 10 mg Ascorbic acid  6 mg

(23) The microgranules were coated with a copolymer of methacrylic acid.

(24) To make the tablets, the microgranules of DAO were compressed with microcrystalline cellulose and magnesium stearate.

Example 3

(25) DAO sachets were prepared containing 100 or 150 mg of DAO microgranules prepared as in the first part of example 1.

Example 4

(26) DAO and caffeine sachets were prepared containing 100 or 150 mg of DAO microgranules prepared as in the first part of example 2.

Example 5

(27) DAO capsules were prepared containing 100 or 150 mg of DAO microgranules prepared as in the first part of example 1, filling the soft gelatin capsules with these microgranules.

Example 6

(28) DAO and caffeine capsules were prepared, containing 100 or 150 mg of the DAO microgranules prepared as in the first part of example 2, filling the soft gelatin capsules with these microgranules.

Example 7

(29) Determination of the efficacy of DAO compositions, the object of the present invention, in children with a diagnosis of attention deficit hyperactivity disorder and who present with DAO deficiency.

(30) The study was carried out with 60 children selected with ages of between 8/10 years to 18 years, diagnosed with attention deficit hyperactivity disorder, as out-patients. Of these 60,45 were boy and 15 girls, since girls often only present symptoms of attention deficit whereas hyperactivity is more common in boys.

(31) Before starting treatment with DAO administration, the children were selected using the results of DAO activity level in plasma. Reduced DAO activity was considered to be between 80 and 40 HDU/ml and very reduced activity was below 40 HDU/ml. Therefore, those children with reduced DAO values, that is, below 80 HDU/ml, were included in the study, although the symptoms of ADHD were most clearly seen in children with DAO activity below 60 HDU/ml. Of the 60 children with ADHD diagnosis participating in the study, 78% showed activity below the threshold of normal (80 HDU/ml) and 39% showed activity below 40, that is had “very reduced” activity. Therefore, of the 100% of children diagnosed, 78% participated in the study whereas the other 22% had ADHD originating in other causes.

(32) Oral compositions containing DAO, alone or associated with caffeine, object of the present invention, were tested in a total of 47 children selected with ages between 8/10 years to 18 years, diagnosed with attention deficit hyperactivity disorder, as out-patients. Of these 47, 36 were boys and 11 girls, and they were randomly assigned to receive compositions of DAO, DAO and caffeine or placebo.

(33) In addition to DAO treatment, milk was removed from the diet of the children participating in the study; this is a food consumed daily and recurrently that contains histamine and other milk proteins that cause endogenous release of histamine, so that ingestion of additional DAO is more efficient if an important source of histamine such as milk is removed at the same time.

(34) Methods based on school performance were used for evaluation of treatment. Children with a diagnosis of ADHD have poor performance in school and, when through treatment they reduce hyperactivity and recover attention, they clearly improve their school results in both academic results and attitude.

(35) The table below was used to quantify the result of treatment: “Child Attention Profile” (CAP) diagnostic evaluation scale that is based on the observation and scoring of 12 items (Cozza S. J. y col., Tratamiento de Niños y Adolescentes, Tomo II, capitulo XXXIII, 1399-1452 en Hales R E, Yudosfky S C (ed), Tratamiento de Psiquiatría Clínica, Barcelona Masson 2004) [Cozza S. J. et al., Treatment of Children and Adolescents, Volume II, Chapter XXXIII, 1399-1452 in Hales R E, Yudosfky S C (ed,), Clinical Psychiatric Treatment, Barcelona Masson 2004]:

(36) TABLE-US-00003 Child Attention Profile (CAP) False Occasionally Frequently 1 Fails to finish things he/she starts 2 Can't concentrate, can't pay attention for long 3 Can't sit still, restless, or hyperactive 4 Fidgets 5 Daydreams or gets lost in their thoughts 6 Impulsive or acts without thinking 7 Difficulty in following directions 8 Talks out of turn 9 Messy work 10 Inattentive, easily distracted 11 Talks too much 12 Fails to carry out assigned tasks The 12 items are scored 0, 1, 2. The total score is the sum of all the items. Sub-scores: Lack of attention (sum of scores of items 1,2,5,7,9,10 and 12); Hyperactivity (sum of scores of items 3,4,6,8,11).

(37) Recommended scores as an upper limit of normality (percentile 93)

(38) TABLE-US-00004 Boys Girls Lack of Attention 9 7 Hyperactivity 6 5 Total score 15 11

(39) The “Abbreviated Conners” questionnaire was also used:

(40) TABLE-US-00005 Not at Just a Pretty Very all little much much 1 Restless or overactive 2 Excitable, impulsive 3 Disturbs other children 4 Fails to finish things he/she starts 5 Constantly fidgeting 6 Inattentive, easily distracted 7 Demands must be met immediately, easily frustrated 8 Cries often and easily 9 Mood changes quickly and drastically 10 Temper outbursts, explosive and unpredictable behaviour

(41) The Connors behavioural questionnaire for parents is a guideline for recording the most significant hyperactive behaviours that may be shown in possible attention disorders, which must be completed by parents, enabling them to also have a degree of clarity into minor problems. It consists of 10 items that must be completed with a score of 0 to 3 points; with 0 corresponding to the absence of the observed item in the person being evaluated and the value 3 for constant and common presence. The maximum score is 30. Between 0 to 10 points: normally active, does not have problems; from 10 to 20 points: situational hyperactivity or normally active but immature in temperament; from 20 to 30 points: very hyperactive or disruptive. In general terms, for boys between 6 and 11 years, ADHD is suspected with a score higher than 16 points. For girls between 6 and 11 years, ADHD is suspected with a score higher than 12 points.

(42) The following tables show the results of reduction of symptoms caused by ADHD after the administration of a dosage protocol of 4 mg DAO twice a day to 37 children diagnosed with ADHD and with DAO deficiency compared to 10 children who were administered placebo.

(43) TABLE-US-00006 TABLE 1 Comparative results of observation and scoring the 12 items of the “Child Attention Profile” (CAP) between children who took DAO tablets, of Example 1, and those who did not take DAO. These results show a clear improvement in the attention profile of treated children, both boys and girls, in comparison to the results obtained after placebo administration. Initial profile Final profile Boys with DAO of Lack of attention 10-14 6-9 Example 1 Hyperactivity  8-10 4-6 Total 18-24 10-15 Boys with placebo Lack of attention 10-14 10-13 Hyperactivity  8-10  7-10 Total 18-24 14-23 Girls with DAO of Lack of attention  9-14 5-7 Example 1 Hyperactivity  7-10 3-5 Total 16-24  8-12 Girls with placebo Lack of attention  9-14  8-14 Hyperactivity  7-10 7-9 Total 16-24 15-23

(44) TABLE-US-00007 TABLE 2 Comparative results of the observation and scoring of the 10 items of the “Abbreviated Conners” questionnaire between children taking DAO tablets, of Example 1, and those that did not take DAO. These results show a clear improvement in the attention profile of treated children, both boys and girls, in comparison to the results obtained after placebo administration. Initial score Final score Boys with DAO of Example 1 16-30  8-15 Girls with DAO of Example 1 12-30  6-12 Boys with placebo 16-30 15-29 Girls with placebo 13-30 12-27

(45) TABLE-US-00008 TABLE 3 Comparative results of the observation and scoring of the 10 items of the “Abbreviated Conners” questionnaire between children taking DAO and caffeine tablets, of Example 2, and those who did not take DAO. These results show a clear improvement in the attention profile of treated children, both boys and girls, in comparison to the results obtained after placebo administration. Initial score Final score Boys with DAO and caffeine of 16-29  7-14 Example 2 Girls with DAO and caffeine of 12-28  5-10 Example 2 Boys with placebo 16-29 15-29 Girls with placebo 13-27 12-27